CN102584965A - Bryophyte reversal-resistant protein PpLEA3-20 and encoding gene and application thereof - Google Patents
Bryophyte reversal-resistant protein PpLEA3-20 and encoding gene and application thereof Download PDFInfo
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Abstract
The invention discloses a bryophyte reversal-resistant protein PpLEA3-20 and an encoding gene and an application thereof. A plant resistance-related protein provided by the invention is named PpLEA3-20, is derived from physcomitrellapatens, and is a protein shown as (1) or (2), wherein (1) is constituted by an amino acid sequence shown as a sequence 2 in a sequence table, or (2) is obtained by performing substitution and/or deletion and/or addition of one or more amino acid residues on the amino acid residue sequence shown as the sequence 2 in the sequence table, is relevant to plant resistance and is derived from (1). As proved by an experiment for introducing the encoding gene of PpLEA3-20 into oryza sativassp.japonica, the survival rates (44 percent, 36 percent, 60 percent and 72 percent) and growth states of four obtained T2-generation PpLEA3-20 transgenic plants (20-1, 20-2, 20-3 and 20-4) in a drought stress experiment are remarkably better than those of a check plant.
Description
Technical field
The present invention relates to biological technical field, the anti-contrary albumen PpLEA3-20 of particularly a kind of bryophyte and encoding sox and application.
Background technology
The factors of coercing such as the arid in the physical environment, saline and alkaline, low temperature have material impact to growth and development of plant, can cause the extensive underproduction of farm crop when serious, and cultivating the resistance of reverse crop is one of major objective of plant husbandry.Current, the crop varieties that has resistance of reverse through the genetic engineering breeding acquisition is a kind of efficient ways.And the technical bottleneck problem of most critical is the screening and the function discovery of effective adversity gene in this method.
Bryophyte is the land pioneer plant that occur the ancient ordovician period before 500,000,000 years, and the gametophyte with " cormus " in its life history is main vegetative growth phase." cormus " blade is made up of monolayer, only locates to be made up of several confluent monolayer cells at its " middle rib ", and the weave construction of regulation and control such as no transfusion tissue and pore water metabolism is keeping the significantly characteristic of waterplant.Owing to lack water conduction and regulator control system, therefore, when original " bryophyte " dried up landing, must at first coerce greatly: water deficit, temperature shock in the face of two.Huge selective pressure forces bryophyte to be evolved out to be different from the adverse circumstance coping mechanism of " vascular plant " (fern, spermatophyte).The fact shows: be present in the more intracellular genes of bryophyte and bearing the responsibility of tackling serious adverse circumstance, they can protect cell to avoid the adverse circumstance injury.
Summary of the invention
The purpose of this invention is to provide a kind of albumen relevant and encoding sox thereof with stress resistance of plant.
The albumen relevant with stress resistance of plant provided by the present invention, name is called PpLEA3-20, derive from small liwan moss (Physcomitrella patens), is following 1) or 2) protein:
1) protein of forming by the aminoacid sequence shown in the sequence in the sequence table 2;
2) with the amino acid residue sequence of sequence in the sequence table 2 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and relevant with stress resistance of plant by 1) deutero-protein.
Sequence table sequence 2 is the aminoacid sequence of PpLEA3-20, comprises 208 amino acid.
In order to make 1) in PpLEA3-20 be convenient to purifying, label as shown in table 1 on proteinic N-terminal that can the aminoacid sequence shown in the sequence 2 is formed in by sequence table or C-terminal connect.
The sequence of table 1. label
| Label | Residue | Sequence |
| Poly-Arg | 5-6 (being generally 5) | RRRRR |
| Poly-His | 2-10 (being generally 6) | HHHHHH |
| FLAG | 8 | DYKDDDDK |
| Strep-tag?II | 8 | WSHPQFEK |
| c- |
10 | EQKLISEEDL |
Above-mentioned 2) but in the PpLEA3-20 synthetic, also can synthesize its encoding sox earlier, carry out biology again and express and to obtain.Above-mentioned 2) encoding sox of the PpLEA3-20 in can be through lacking sequence in the sequence table 1 codon of one or several amino-acid residue in the dna sequence dna shown in the 5 ' terminal 118-744 bit base; And/or carry out the missense mutation of one or several base pair, and/or obtain at the encoding sequence that its 5 ' end and/or 3 ' end connects the label shown in the table 1.
Above-mentioned and encoding sox plant adversity resistance related protein also belongs to protection scope of the present invention.
Specifically can be following 1 with the encoding sox of plant adversity resistance related protein)-4) in arbitrary described gene:
1) its encoding sequence is the deoxyribonucleotide from 5 ' terminal 118-744 position of sequence 1 in the sequence table;
2) its nucleotide sequence is the sequence 1 in the sequence table;
3) under stringent condition with 1) or 2) gene recombination and the gene of encoding said proteins;
4) with 1) or 2) gene have homology and the gene of encoding said proteins more than 90%.
Above-mentioned stringent condition can be that (or 0.1 * SSC), the solution of 0.1%SDS is hybridized under 65 ℃ and washed film with 0.1 * SSPE in DNA or RNA hybrid experiment.
Increase above-mentioned PpLEA3-20 full length gene or its arbitrary segmental primer to also belonging to protection scope of the present invention.
Contain above-mentioned and expression cassette, recombinant vectors, transgenic cell line and reorganization bacterium the plant adversity resistance related protein encoding sox and also belong to protection scope of the present invention.
Available existing plant expression vector construction contains the recombinant expression vector of PpLEA3-20 gene.Said plant expression vector comprises double base agrobacterium vector and the carrier etc. that can be used for the plant micropellet bombardment, like pCAMBIA3301, pCAMBIA1300, pBI121, pBin19, pCAMBIA2301, pCAMBIA1301-UbiN or other plant expression vector of deriving.
When using the gene constructed recombinant expression vector of PpLEA3-20; Can before its transcription initiation Nucleotide, add any enhancement type, composing type, organizing specific type or inducible promoter; Like cauliflower mosaic virus (CAMV) 35S promoter, ubiquitin (Ubiquitin) gene promoter (pUbi) etc., they can use separately or be used in combination with other plant promoter; In addition; When using gene constructed plant expression vector of the present invention; Also can use enhanser, comprise translational enhancer or transcriptional enhancer, these enhanser zones can be ATG initiator codon or neighboring region initiator codon etc.; But must be identical with the reading frame of encoding sequence, to guarantee the correct translation of whole sequence.The source of said translation wave and initiator codon is widely, can be natural, also can be synthetic.Translation initiation region can be from transcription initiation zone or structure gene.
For the ease of transgenic plant cells or plant being identified and screening; Can process used plant expression vector, as be added in the plant to express and to produce the enzyme of colour-change or the gene of luminophor (gus gene, GFP gene, luciferase genes etc.), have antibiotic marker thing (qingfengmeisu qiong affinity tag, kantlex affinity tag etc.) or the anti-chemical reagent marker gene (like anti-weedkiller gene) of resistance etc.From the security consideration of transgenic plant, can not add any selected marker, directly with adverse circumstance screening transformed plant.
Said recombinant expression vector specifically can be the recombinant expression vector that insertion is above-mentioned and encoding sox plant adversity resistance related protein obtains between the MCS of the plasmid pC1390 after inserting corn Ubiquitin promotor.
Another object of the present invention provides a kind of method of cultivating the transgenic plant of resistance raising.
The method of the transgenic plant that cultivation resistance provided by the present invention improves is that above-mentioned encoding sox PpLEA3-20 with plant adversity resistance related protein is imported in the purpose plant, obtains the transgenic plant that resistance is higher than the purpose plant.
Said encoding sox PpLEA3-20 with plant adversity resistance related protein imports in the plant through said recombinant expression vector.
Above-mentioned resistance specifically can be drought tolerance.
Conventional biological methods such as the plant expression vector that carries of the present invention and plant adversity resistance related protein encoding sox PpLEA3-20 can lead through Ti-plasmids, Ri plasmid, plant viral vector, directly DNA conversion, microinjection, electricity, agriculture bacillus mediated are transformed in the plant.Said plant is dicotyledons or monocotyledons, preferably paddy rice.
The present invention can obtain arid tolerance enhanced transgenic cell line and transfer-gen plant.
The present invention has found that through multiple molecular biology research technology a gene and expressing protein PpLEA3-20 thereof have significant degeneration-resistant border ability from bryophyte.PpLEA3-20 is a member in Late Embryogenesis Abundant protein (LEA) family.LEA gene after deliberation all plays important effect in different vital processes, the key protein molecule when being organism antagonism environment-stress.
The experiment that PpLEA3-20 gene of the present invention is imported wild-type paddy rice (Oryza sativa ssp.japonica) can prove: T
24 of generation are changeed PpLEA3-20 gene plant (20-1,20-2,20-3,20-4), and in the drought stress experiment, survival rate (44%, 36%, 60% and 72%) and growth conditions all will obviously be better than wild-type contrast and empty carrier adjoining tree.Explain that PpLEA3-20 gene provided by the invention and its encoded protein can significantly improve the drought tolerance of plant.To cultivating the plant with adverse resistance kind; The new variety such as crop, woods grass of particularly cultivating anti-abiotic stress (drought-enduring and/or salt tolerant) have important theory and practical significance, can be used for the cultivation and the evaluation of the required resistance of reverse plant variety of husbandry and ecological environment treatment.
Description of drawings
Fig. 1 is the proteinic two dimensional gel electrophoresis collection of illustrative plates of small liwan moss cormus, and arrow indication PpLEA3-20 albumen abundance in the plant that arid is handled significantly increases.The adjoining tree of A. handling without arid; B. arid is handled 3 days plant.
Fig. 2 is the structure of plant expression vector pCU-PpLEA3-20.
Fig. 3 identifies that for the PCR of the rice plant of commentaries on classics PpLEA3-20 gene WT is an adjoining tree among the figure; 20-1,20-2,20-3,20-4 are 4 transgenic paddy rice strain systems.
Fig. 4 is the expression analysis of PpLEA3-20 gene in rice plant, and A. changes the two dimensional gel electrophoresis collection of illustrative plates of PpLEA3-20 trans-genetic hybrid rice plant among the figure, arrow indication PpLEA3-20 expression of gene product, a. adjoining tree, b. transfer-gen plant; The PMF collection of illustrative plates of B.PpLEA3-20 gene expression product; The CID collection of illustrative plates of C.PpLEA3-20 gene expression product; The complete sequence of D.PpLEA3-20 genetic expression albumen PpLEA3-20, the peptide section sequence that the CID collection of illustrative plates identifies is represented with black matrix.
Fig. 5 is for showing that commentaries on classics PpLEA3-20 trans-genetic hybrid rice plant and adjoining tree are at the plant height of growth in the time of 45 days.
Fig. 6 changes PpLEA3-20 trans-genetic hybrid rice plant and the survival rate of control plant after drought stress is handled for showing.
Fig. 7 is for changeing the experiment of PpLLEA3-20 gene plant drought tolerance; Adjoining tree (WT) and 4 transgenic lines (20-1,20-2,20-3; 20-4) arid is handled rehydration after 10 days under the same conditions; Be illustrated as the plant of rehydration after 15 days, wherein the wild-type contrast is consistent with the experimental result phenotype of empty carrier adjoining tree, so only show a kind of result of contrast.
Embodiment
Below in conjunction with specific embodiment the present invention is described further, but the present invention is not limited to following examples.
Among the following embodiment,, be ordinary method like no specified otherwise.
The screening of embodiment 1, liver moss stress tolerance correlative protein PpLEA3-20 encoding sox PpLEA3-20 and cDNA clone thereof.
The small liwan moss arid is handled: will be grown in small liwan moss (Physcomitrella patens) (Geng Xuke etc., 2008 plant genes target practice---the application of model plant small liwan moss on the BCD substratum.The biology circular, 43 (4): 13-15; The public can obtain from Capital Normal University) cormus is transferred on the wetted filter paper, puts into the 500ml beaker, seal with thieving paper.24 ± 1 ℃ of temperature, carry out arid in the culturing room of relative humidity 30 ± 5% and handle.After 3 days, the cormus that takes out arid is used for proteins extraction.Get the cormus of not arid processing and do control experiment.
The method that the proteins extraction of small liwan moss cormus adopts the substep extracting to combine with the phenol extracting.From under the normal condition with drought condition under protein separate with fluorescence difference gel electrophoresis (DIGE) through classical IEF/SDS-PAGE two-dimensional electrophoresis (2DE).Concrete steps are carried out according to the experimental implementation guide that Amersham Bioscience provides.
The protein pattern that obtains passes through ImageMaster 2D Platinum (Amersham Bioscience) and DeCyder 2D Software respectively, and Version 6.5 (Amersham Bioscience) analyzes.Abundance changes the separated acquisition of protein spots above 2 times under drought condition.All differential protein spots are used trypsin treatment respectively, and institute's polypeptide that obtains is identified with mass spectrograph (MALDI TOF/TOF MS) and DB (NCBI).We find, under drought condition, have a protein on the two-dimensional electrophoresis (2DE) of classics and fluorescence difference gel electrophoresis (DIGE) gel, raised respectively 4.45 times and 5.99 times (Figure 1A, B).This protein is named as PpLEA3-20.
Identify the sequence that obtains according to mass spectrum, design primer, the cDNA sequence of acquisition PpLEA3-20 gene.Concrete operations are following: at first by above-mentioned condition the small liwan moss cormus is implemented arid and handle to induce the PpLEA3-20 expression of gene; Then, extract the total RNA of small liwan moss cormus, RNA is synthesized cDNA with reversed transcriptive enzyme.According to the mass spectrum qualification result.
The design primer is following: 5 '-GGGGTACCCCTTCCCACTAAATACCGT-3 ' and 5 '-GGACTAGTTCCAGAGCCAATGCTTG-3 '.
The cDNA that obtains with reverse transcription is a template, carries out pcr amplification, the PCR product is carried out 0.8% agarose gel electrophoresis detect, and obtains the band that molecular weight is about 829bp, conforms to expected results.Reclaim test kit (TIANGEN) with sepharose and reclaim this fragment.Should reclaim fragment is connected with pMD19-T Simple (Takara); To connect product transformed into escherichia coli DH5 α competent cell; According to the carboxylic Bian penicillin resistance label screening positive colony on the pMD19-T Simple carrier, obtain containing the segmental recombinant plasmid of recovery.Universal primer M13 with on this recombinant plasmid vector carries out nucleotide sequencing to it; Sequencing result shows that the gene that increases (name PpLEA3-20) is made up of 829 deoxyribonucleotides; Its ORFs (ORF) for sequence 1 in the sequence table from 5 ' terminal the 118th to 744 deoxyribonucleotide, encoding amino acid sequence is the protein of sequence 2 in the sequence table.
Embodiment 2, commentaries on classics PpLEA3-20 gene are cultivated drought-enduring plant and check and analysis thereof
1) structure of PpLEA3-20 expression vector pCU-PpLEA3-20
Just in the sequence table corn Ubiquitin promotor shown in the sequence 3 (with 5 '-CCAAGCTTAAGCTTTCTAGTGCAGTGCAGCGTGAC-3 ' and 5 '-GGGGTACCCTGCAGCCTCTAGTGCAGAAGTAACACCA-3 ' is primer; From corn gene group DNA, increase and obtain) insert plant binary expression vector pC1390 (available from Cambia; Queenslan d.Australia) in HindIII and the Kpn I site, constitutes intermediate carrier pC1390-Ubi.The cDNA fragment of the PpLEA3-20 gene that obtains increasing in the foregoing description 1 is with Spe I and Kpn I double digestion, and forward obtains recombinant vectors after inserting the Ubiquitin promotor of this intermediate carrier pC1390-Ubi.The recombinant expression vector called after pCU-PpLEA3-20 (as shown in Figure 2) of the PpLEA3-20 gene of deoxyribonucleotide from 5 ' terminal 1-829 position that will contain sequence 1 in the ordered list.This carrier has Ka Na and two selection markers of Totomycin.
2) acquisition and the detection of changeing the PpLEA3-20 gene plant are identified
The mode of pCU-PpLEA3-20 carrier with electric shock changed among the Agrobacterium LBA4404.The picking positive colony is in 28 ℃ of cultivations.Treat that it is between 0.120～0.140 the time that Agrobacterium concentration reaches the OD600 value, infect paddy rice (Oryza sativa ssp.japonica) callus, infect 30min with this bacterium liquid, during shake with have gentle hands frequently.
Then callus is transferred on the common culture medium, lucifuge is cultivated 3～4d altogether.Cultivate altogether finish after, callus transferred to selects to cultivate on the substratum 10～15d follow-up generation once.New HYG (Totomycin) resistant calli that picking comes out from former callus surface growth is transferred on the presorting substratum, presorting cultivation 7d under 28 ℃ of dark.Select creamy white, ganoid callus is transferred on the division culture medium, 28 ℃ of differentiation culture: under dark, cultivate 3d earlier, under lasting cold light shines, cultivate 15～20d then.To transfer on the root induction substratum from the seedling that resistant calli bears again, continuous light is cultivated 15d, and the seedling that upgrowth situation is good is directly transferred to field planting.Gather in the crops transgenic paddy rice seed then autumn.
After the seed results, be sowed on the MS screening culture medium that contains Totomycin (30mg/L) and screen.Treat that T1 moves on in the land for growing field crops when growing to the 4-6 leaf for plant and grow.For after the individual plant results, each single-strain seed is sowed respectively with T1, continues screening to observe the T2 separation case in generation with Totomycin, and so repeat number generation is until obtaining stable commentaries on classics PpLEA3-20 strain system.
Select 4 high strains systems of PpLEA3-20 genetic expression in the commentaries on classics PpLEA3-20 strain system of the genetic stability that above-mentioned screening obtains, carry out PCR, protein two dimensional gel electrophoresis and mass spectrum checking respectively.
PCR checking: extract oryza sativa genomic dna with the CTAB method, use forward primer (5 '-CCTTCCCACTAAATA CCGT-3 ') and reverse primer (5 '-TCCAGAGCCAATGCTTG-3 ') carry out pcr amplification.All can amplify the purpose band (as shown in Figure 3) of 829bp in above-mentioned 4 strains system.
The checking of protein two dimensional gel electrophoresis: using classification extraction process (Cui etc., 2005) to extract earlier changes the whole protein of PpLEA3-20 trans-genetic hybrid rice plant and wild-type contrast rice plant.Obtain protein two dimensional gel electrophoresis collection of illustrative plates by ordinary method.Comparative analysis is found, in changeing PpLEA3-20 trans-genetic hybrid rice plant, possible PpLEA3-20 gene expression product (shown in A among Fig. 4) occurs.
The mass spectrum checking: the possible PpLEA3-20 gene expression product in the above-mentioned cutting-out gel, use trypsin digestion.Identify through MALDI TOF/TOF mass spectrum, confirm this protein PpLEA3-20 expression of gene albumen (shown in B, C, D among Fig. 4) just.
The drought tolerance of 3) changeing PpLEA3-20 gene strain system is identified
With in the empty carrier pC1390-Ubi importing wild-type paddy rice, prepare the empty carrier contrast according to the method that obtains to change PpLEA3-20 gene strain system.
This step is provided with empty carrier contrast and wild-type contrast.
With above-mentioned be tested and appraised to confirm as change 4 of PpLEA3-20 trans-genetic hybrid rice strains system (20-1,20-2,20-3 and 20-4) and adjoining trees (empty carrier contrast and wild-type contrast) and plant respectively in 25 hole basins.Developing medium is commercially available nutrition soil, and culture temperature is 26 ± 1 ℃.Fully growing 45 days under the water conditions.Change the extraordinary growth consistence that the PpLEA3-20 plant shows, but the plant height comparison reduces about 8.1% (as shown in Figure 5, the empty carrier contrast is consistent with wild-type adjoining tree phenotype, has omitted the empty carrier map) according to plant; Carrying out arid handles: stop to supply water, treating that rice leaf is accomplished curls and when wilting, resumes water supply.After resuming water supply 15 days, observing not respectively, homophyletic is the survival condition of paddy rice and takes a picture (like Fig. 6, shown in Figure 7, the empty carrier contrast is consistent with wild-type adjoining tree phenotype, has omitted the empty carrier map).
Three repetitions are established in experiment altogether, and in repeating, each strain of being tested is that plant is respectively 25 strains at every turn.The result shows: adjoining tree is after arid processing and rehydration, and survival rate is 0, and is promptly dead fully under above-mentioned arid treatment condition; And 4 survival rates of changeing the strain system of PpLEA3-20 gene are respectively 44%, 36%, 60% and 72% (Fig. 6).Explain that the drought tolerance of changeing the PpLEA3-20 gene plant obviously is superior to adjoining tree.
Among the land for growing field crops, it is ripe until seed to observe its development condition with the commentaries on classics PpLEA3-20 trans-genetic hybrid rice plant kind of surviving after the above-mentioned arid processing.The result finds: the rice plant that changes the PpLEA3-20 gene does not have significant difference with adjoining tree in its strain shape, heading aspect period, but plant height descends about 10 centimetres.
Claims (10)
1. an albumen is following 1) or 2) protein:
1) protein of forming by the aminoacid sequence shown in the sequence in the sequence table 2;
2) with the amino acid residue sequence of sequence in the sequence table 2 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and relevant with stress resistance of plant by 1) deutero-protein.
2. the said proteic encoding sox of claim 1.
3. encoding sox according to claim 2 is characterized in that: said proteic encoding sox is following 1)-4) in arbitrary described gene:
1) its encoding sequence is the deoxyribonucleotide from 5 ' terminal 118-744 position of sequence 1 in the sequence table;
2) its nucleotide sequence is the sequence 1 in the sequence table;
3) under stringent condition with 1) or 2) gene recombination and the said proteic gene of coding claim 1;
4) with 1) or 2) gene have the homology 90% or more and the said proteic gene of claim 1 of encoding.
4. contain claim 2 or 3 said expression of gene boxes, transgenic cell line or reorganization bacterium.
5. the recombinant vectors that contains claim 2 or 3 said genes.
6. recombinant vectors according to claim 5 is characterized in that: insert the recombinant expression vector that claim 2 or 3 said genes obtain between the MCS of said recombinant vectors for the plasmid pC1390 after inserting corn Ubiquitin promotor.
7. total length or its arbitrary segmental primer of amplification claim 2 or 3 said genes are right.
8. a method of cultivating transgenic plant is that claim 2 or 3 described encoding soxs are changed in the purpose plant, obtains the resistant transgenic plant.
9. method according to claim 8 is characterized in that: claim 2 or 3 described encoding soxs are to import in the plant through claim 5 or 6 described recombinant expression vectors.
10. according to Claim 8 or 9 described methods, it is characterized in that: said resistant transgenic plant is drought-enduring transgenic plant; Said purpose plant is dicotyledons or monocotyledons, preferably paddy rice.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102659936A (en) * | 2012-04-12 | 2012-09-12 | 中国农业科学院生物技术研究所 | Plant-stress-tolerance related protein, its encoding gene and application |
| CN102776229A (en) * | 2012-07-26 | 2012-11-14 | 首都师范大学 | Moss PpDHN-31 protein and application of coding gene thereof for improving drought resistance of plant |
| CN102964438A (en) * | 2012-11-29 | 2013-03-13 | 中国农业科学院生物技术研究所 | Stress-resistance-related protein PpLEA3-23 of plant as well as coding gene and application of protein |
| CN103911385A (en) * | 2014-04-24 | 2014-07-09 | 中国科学院西北高原生物研究所 | Medicago ruthenica late-embryogensis abundant protein gene and application thereof |
| CN104726488A (en) * | 2015-03-29 | 2015-06-24 | 河北科技大学 | Method for culturing stress-resistance herbicide-resistance transgenic aerobic rice |
| CN105801679A (en) * | 2016-04-15 | 2016-07-27 | 首都师范大学 | Application of protein PpLEA3-2 in regulating and controlling plant stress resistance |
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2011
- 2011-01-10 CN CN201110004413.3A patent/CN102584965B/en not_active Expired - Fee Related
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102659936A (en) * | 2012-04-12 | 2012-09-12 | 中国农业科学院生物技术研究所 | Plant-stress-tolerance related protein, its encoding gene and application |
| CN102776229A (en) * | 2012-07-26 | 2012-11-14 | 首都师范大学 | Moss PpDHN-31 protein and application of coding gene thereof for improving drought resistance of plant |
| CN102776229B (en) * | 2012-07-26 | 2013-09-04 | 首都师范大学 | Moss PpDHN-31 protein and application of coding gene thereof for improving drought resistance of plant |
| CN102964438A (en) * | 2012-11-29 | 2013-03-13 | 中国农业科学院生物技术研究所 | Stress-resistance-related protein PpLEA3-23 of plant as well as coding gene and application of protein |
| CN102964438B (en) * | 2012-11-29 | 2014-05-07 | 中国农业科学院生物技术研究所 | Stress-resistance-related protein PpLEA3-23 of plant as well as coding gene and application of protein |
| CN103911385A (en) * | 2014-04-24 | 2014-07-09 | 中国科学院西北高原生物研究所 | Medicago ruthenica late-embryogensis abundant protein gene and application thereof |
| CN103911385B (en) * | 2014-04-24 | 2016-03-30 | 中国科学院西北高原生物研究所 | Medicago ruthenica late embryo generation Abundant protein gene and application thereof |
| CN104726488A (en) * | 2015-03-29 | 2015-06-24 | 河北科技大学 | Method for culturing stress-resistance herbicide-resistance transgenic aerobic rice |
| CN105801679A (en) * | 2016-04-15 | 2016-07-27 | 首都师范大学 | Application of protein PpLEA3-2 in regulating and controlling plant stress resistance |
| CN105801679B (en) * | 2016-04-15 | 2019-04-09 | 首都师范大学 | Application of the protein PpLEA3-2 in regulation stress resistance of plant |
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